Quantitative X-ray microanalysis of hydrogen peroxide within plant cells

✍ Scribed by Shaoliang Chen; Andrea Olbrich; Rosemarie Langenfeld-Heyser; Eberhard Fritz; Andrea Polle

• Publisher: John Wiley and Sons
• Year: 2009
• Tongue: English
• Weight: 595 KB
• Volume: 72
• Category: Article
• ISSN: 1059-910X
• DOI: 10.1002/jemt.20639

✦ Synopsis

Abstract

Using quantitative X‐ray microanalysis in combination with CeCl~3~‐based cytochemical staining of hydrogen peroxide (H~2~O~2~) we have developed a new solution for quantification of H~2~O~2~ at the subcellular level. Quantitative X‐ray microanalysis of plastic‐embedded leaves of Populus euphratica Oliv. showed that the obtained cerium precipitates by CeCl~3~ staining were the mixture of cerium perhydroxides and cerium phosphate, in which the fractions of CePO~4~ were: (1) 52–74% in cell walls of fresh leaf segments, and (2) 34–70% in the cytoplasm in 10 mM H~2~O~2~‐treated leaf segments that were previously freeze‐dried. Taking the concentration of cerium phosphate as staining background, we reached the cellular concentration of cerium perhydroxides and the corresponding concentration of H~2~O~2~. Results showed that H~2~O~2~ was present in the cytoplasm of rehydrated leaf segments (29–58 mM), but in fresh leaves, H~2~O~2~ was observed in the walls of all measured cell types (17–74 mM). Microsc. Res. Tech., 2009. © 2008 Wiley‐Liss, Inc.